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Galactosylation reactions catalyzed by disaccharide phosphorylases

Chao Chen (UGent)
(2012)
Author
Promoter
(UGent) and (UGent)
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Abstract
Galactosylation, as one of the main types of glycosylation reactions, is of particular importance both in vivo and in vitro. The goal of this PhD research is to explore and exploit galactosylation reactions catalyzed by disaccharide phosphorylases. On the one hand, β-galactoside phosphorylases from the novel family GH-112 were evaluated for their ability to synthesize β-galactosides. Mutational analysis of their acceptor specificity was performed, and (ga)lacto-N-biose phosphorylase was combined with lactose phosphorylase in a cascade reaction to produce bifidogenic disaccharides. On the other hand, the trehalose phosphorylase from Caldanaerobacter subterraneus (CsTP) was targeted to produce α-galactosides. More specifically, lactotrehalose was synthesized by exploiting its promiscuous activity, and the product was purified using activated carbon. Furthermore, the enzyme’s donor specificity was modified from glucose to galactose by means of iterative saturation mutagenesis, thereby producing β-galactose-1-phosphate (βGal-1P) from lactotrehalose. Based on these results, we propose the engineering of trehalose phosphorylase as a general strategy for the production of glycosyl phosphates, making use of the special features of trehalose as substrate.
Keywords
directed evolution, sugar phosphate, donor specificity, trehalose phosphorylase, galactosylation, (ga)lacto-N-biose phosphorylase

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MLA
Chen, Chao. Galactosylation Reactions Catalyzed by Disaccharide Phosphorylases. Ghent University. Faculty of Bioscience Engineering, 2012.
APA
Chen, C. (2012). Galactosylation reactions catalyzed by disaccharide phosphorylases. Ghent University. Faculty of Bioscience Engineering, Ghent, Belgium.
Chicago author-date
Chen, Chao. 2012. “Galactosylation Reactions Catalyzed by Disaccharide Phosphorylases.” Ghent, Belgium: Ghent University. Faculty of Bioscience Engineering.
Chicago author-date (all authors)
Chen, Chao. 2012. “Galactosylation Reactions Catalyzed by Disaccharide Phosphorylases.” Ghent, Belgium: Ghent University. Faculty of Bioscience Engineering.
Vancouver
1.
Chen C. Galactosylation reactions catalyzed by disaccharide phosphorylases. [Ghent, Belgium]: Ghent University. Faculty of Bioscience Engineering; 2012.
IEEE
[1]
C. Chen, “Galactosylation reactions catalyzed by disaccharide phosphorylases,” Ghent University. Faculty of Bioscience Engineering, Ghent, Belgium, 2012.
@phdthesis{3009057,
  abstract     = {Galactosylation, as one of the main types of glycosylation reactions, is of particular importance both in vivo and in vitro. The goal of this PhD research is to explore and exploit galactosylation reactions catalyzed by disaccharide phosphorylases. On the one hand, β-galactoside phosphorylases from the novel family GH-112 were evaluated for their ability to synthesize β-galactosides. Mutational analysis of their acceptor specificity was performed, and (ga)lacto-N-biose phosphorylase was combined with lactose phosphorylase in a cascade reaction to produce bifidogenic disaccharides. On the other hand, the trehalose phosphorylase from Caldanaerobacter subterraneus (CsTP) was targeted to produce α-galactosides. More specifically, lactotrehalose was synthesized by exploiting its promiscuous activity, and the product was purified using activated carbon. Furthermore, the enzyme’s donor specificity was modified from glucose to galactose by means of iterative saturation mutagenesis, thereby producing β-galactose-1-phosphate (βGal-1P) from lactotrehalose. Based on these results, we propose the engineering of trehalose phosphorylase as a general strategy for the production of glycosyl phosphates, making use of the special features of trehalose as substrate.},
  author       = {Chen, Chao},
  isbn         = {9789059895553},
  keywords     = {directed evolution,sugar phosphate,donor specificity,trehalose phosphorylase,galactosylation,(ga)lacto-N-biose phosphorylase},
  language     = {eng},
  pages        = {219},
  publisher    = {Ghent University. Faculty of Bioscience Engineering},
  school       = {Ghent University},
  title        = {Galactosylation reactions catalyzed by disaccharide phosphorylases},
  year         = {2012},
}